LOCUS       sequence-330606-        2964 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB5 entry clone for mammalian expression using the 
            murine RNA polymerase 2 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2964)
  AUTHORS   Wall VE, Garvey LA, Mehalko JL, Procter LV, Esposito D
  TITLE     Combinatorial assembly of clone libraries using site-specific 
            recombination.
  JOURNAL   Methods Mol Biol. 2014;1116:193-208. doi: 
            10.1007/978-1-62703-764-8_14.
  PUBMED    24395366
REFERENCE   2  (bases 1 to 2964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2014"; volume: "1116:193-208. doi"; pages: " 
            10.1007/978-1-62703-764-8_14"
COMMENT     Sequence Label: C453-E25: attB4-mPol2p>-attB5
FEATURES             Location/Qualifiers
     source          1..2964
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..102
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(818..913)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      961..1047
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1139..1166
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1301..1889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1381..1400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2032..2841)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2755..2774
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2833..2852
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata
       61 agcaatgctt ttttataatg ccaactttgt atacaaaagt tggcatcctg aacggcagag
      121 gttacggcag tttgtctctc ccccttccgg gagccacctt cttctccaac cgtcccggtc
      181 gcgctctcgg cgcttctgag gagagaactg gctgagtgac gccctttata gattcgccct
      241 tgtgtcccgc cccttccttt cccgccctcc cttgcgctac ggggccgccc gcaccggcct
      301 acacggagcg cgcgcggcgg agttgttgac gctagggctc cggctccctg gttgggtgtt
      361 ctttctgacg cgacaggagg aggagaatgt tcctggtcct gtcgtcctcc tttcgggttt
      421 cccgtgcact caaaccgagg acttacagaa cggaggataa agttaggcca tttttactca
      481 gcttcggagt tcaggctcat tttcagctaa agtctctcat tagtatcccc ccacacacat
      541 cgggaaaatg gtttgtccta cgcatcggta atgaaggcgg ggcccttcgg gtcctccgga
      601 gcgggttccg ggggtggggg gaaggaggga gggacgggac gggcctcgtt catgaatatt
      661 cagttcaccg ctgaatatgc ataaggcagg caagatggcg cgtccaatca attggaagta
      721 gccgttatta gtggagaggc cccaggacgt tggggcaccg cctgtgctct agtagcttta
      781 cggagccctg gcgctcgatg ttcaagccca agcttgccaa cttttctata caaagttggc
      841 attataaaaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
      901 aaaatcatta tttggagctc catggtagcg ttaacgcgag agtagggaac tgccaggcat
      961 caaataaaac gaaaggctca gtcggaagac tgggcctttc gttttatctg ttgtttgtcg
     1021 gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgtgaagcaa
     1081 cggcccggag ggtggcgggc aggacgcccg ccataaactg ccaggcatca aactaagcag
     1141 aaggccatcc tgacggatgg cctttttgcg tttctacaaa ctcttcctgg ctagcggtaa
     1201 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1321 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1741 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagcttg cgccgtcccg
     1981 tcaagtcagc gtaatgctct gccggtgtta caaccaatta accaattctg attagaaaaa
     2041 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     2101 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     2161 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     2221 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     2281 cgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     2341 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     2401 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     2461 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     2521 tacctggaat gctgtttttc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     2581 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     2641 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     2701 cgcatcgggc ttcccataca agcgatagat tgtcgcacct gattgcccga cattatcgcg
     2761 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgacgt
     2821 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     2881 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     2941 gggccctgcg ccctgcagct ctag
//
